PROJECT SUMMARY: High-Parameter Cytometry Shared Resource (HPCSR) Cytometry is an essential and broadly applicable technology for cancer research to assist with elucidating mechanisms of tumor suppressor and oncogenes, cell-cycle progression, transforming viruses, and to evaluate cancer therapies. It is integral to studies of cancer stem cells, angiogenesis, inflammation, transcriptional regulation in tumor cells, and mechanisms of DNA break and repair. The goal of the High-Parameter Cytometry Shared Resource (HPCSR) is to provide a wide range of Cancer Center investigators with access to cost- effective state-of-the-art instrumentation, expertise and training for their cell sorting, and analysis needs. This technology continues to develop at a rapid pace, especially with the advent of novel tools and reagents, increased computational capacity, and more cost-effective equipment. To keep pace with changing technologies, increased utilization, and growth of users over the past 4 years, the HPCSR has expanded its space with new lab renovations, acquired substantial upgrades of existing equipment, and acquired new major equipment utilizing mass, imaging, high-parameter, and large and small particle cytometry. The main facility is housed in centrally located space and is available to trained users 24 hours a day, 7 days a week by key-card access. Current major instrumentation includes a mass cytometer and imaging system, high-parameter fluorescent cytometer, an imaging cytometer, 4 florescence-activated cell sorters, 7 flow analyzers, and magnetic and large particle cell sorters. The Resource is directed by Dr. Christine Beeton and Mr. Joel Sederstrom, whom both have over 20 years of experience in flow cytometry and is staffed by 5 full-time experienced flow cytometrists who perform operator-assisted cytometry and assist users with data analysis. To ensure optimal use of services, HPCSR provides consultations, group and individually tailored training, and protocols for sample preparation, flow analysis, and cell sorting. This is a heavily used Shared Resource that provided service for 119 Cancer Center investigators during the last year, a substantial increase from 87 users in the first year of the CCSG award. It serves all Programs of the DLDCCC with the Programs in Cell Signaling and Metabolism (CSM), Nuclear Receptor, Transcription and Chromatin Biology (NRTCB), Breast Cancer (BCP), Mechanisms in Cancer Evolution (MCEP), and Pediatric Cancer (PCP) being the biggest users of this Shared Resource. This use translates into an average of 50 publications annually. Future plans include development of new technologies and instrumentation for mass, high-parameter, imaging, large and small particle, and spectral cytometry that are pushing the boundaries of conventional cytometry beyond 50 parameters. Further expansion of space and research staff are required for these new technologies and anticipated continued growth of HPCSR.